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Challenges in Laboratory Detection of Fungal Pathogens in the Airways of Cystic Fibrosis Patients

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Abstract

Study of the clinical significance of fungal colonization/infection in the airways of cystic fibrosis (CF) patients, especially by filamentous fungi, is challenged by the absence of standardized methodology for the detection and identification of an ever-broadening range of fungal pathogens. Culture-based methods remain the cornerstone diagnostic approaches, but current methods used in many clinical laboratories are insensitive and unstandardized, rendering comparative studies unfeasible. Guidelines for standardized processing of respiratory specimens and for their culture are urgently needed and should include recommendations for specific processing procedures, inoculum density, culture media, incubation temperature and duration of culture. Molecular techniques to detect fungi directly from clinical specimens include panfungal PCR assays, multiplex or pathogen-directed assays, real-time PCR, isothermal methods and probe-based assays. In general, these are used to complement culture. Fungal identification by DNA sequencing methods is often required to identify cultured isolates, but matrix-assisted laser desorption/ionization time-of-flight mass spectrometry is increasingly used as an alternative to DNA sequencing. Genotyping of isolates is undertaken to investigate relatedness between isolates, to pinpoint the infection source and to study the population structure. Methods range from PCR fingerprinting and amplified fragment length polymorphism analysis, to short tandem repeat typing, multilocus sequencing typing (MLST) and whole genome sequencing (WGS). MLST is the current preferred method, whilst WGS offers best case resolution but currently is understudied.

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Acknowledgements

CHP is supported by the Midlands Asthma and Allergy Research Association (MAARA) and the National Institute for Health Research (NIHR) Leicester Respiratory Biomedical Research Unit. WM and SC-AC are supported by the National Health and Medical Research Council of Australia and the NSW government. The views expressed here are those of the authors.

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  1. Centre for Infectious Diseases and Microbiology Laboratory Services, ICPMR – Pathology West, Westmead Hospital, 3rd Level ICPMR Building, Westmead, NSW, 2145, Australia

    Sharon C.-A. Chen

  2. Molecular Mycology Research Laboratory, Centre for Infectious Diseases and Microbiology, Sydney Medical School, The University of Sydney, Westmead Hospital, Westmead Institute for Medical Research, Westmead, NSW, Australia

    Sharon C.-A. Chen & Wieland Meyer

  3. Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, Sydney, NSW, Australia

    Wieland Meyer

  4. Department of Infection, Immunity and Inflammation, Institute for Lung Health, University of Leicester, Leicester, LE1 9HN, UK

    Catherine H. Pashley

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Chen, S.CA., Meyer, W. & Pashley, C.H. Challenges in Laboratory Detection of Fungal Pathogens in the Airways of Cystic Fibrosis Patients. Mycopathologia 183, 89–100 (2018). https://doi.org/10.1007/s11046-017-0150-8

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